An Alternative Strategy for Pan-acetyl-lysine Antibody Generation.

An Alternative Strategy for Pan-acetyl-lysine Antibody Generation.

Blog Article

Lysine acetylation is an important post-translational modification in cell signaling.In acetylome studies, a high-quality pan-acetyl-lysine antibody is key to successful enrichment of acetylated peptides for subsequent mass spectrometry analysis.Here we show an alternative method to generate polyclonal pan-acetyl-lysine antibodies using a synthesized random library of acetylated peptides as the antigen.

Our antibodies are tested to be specific for acetyl-lysine peptides/proteins via ELISA and Sailing Vests dot blot.When pooled, five of our antibodies show broad reactivity to acetyl-lysine peptides, complementing a commercial antibody in terms of peptide coverage.The consensus sequence of peptides bound by our antibody cocktail differs slightly from that of the commercial antibody.

Lastly, our antibodies are tested in a proof-of-concept to analyze the acetylome of HEK293 cells.In total we identified 1557 acetylated peptides from 416 proteins.We thus demonstrated that our antibodies are well-qualified for acetylome studies and can complement existing Nail Buffer commercial antibodies.